Production of bi-specific monoclonal antibodies in a hollow-fibre bioreactor

J Immunol Methods. 1993 May 26;161(2):145-50. doi: 10.1016/0022-1759(93)90289-j.

Abstract

In the present study we have measured the feasibility of producing high amounts of bi-specific monoclonal antibodies (MAb) for therapeutic purposes using a hollow-fibre bioreactor. We have studied the isotype composition and functional capacity of an OKT3/OV-TL 3 quadroma (IgG2a/IgG1) and we have compared the production of bi-isotypic MAb in this system with tissue culture flasks or mouse ascites. Both culture supernatant and purified bi-isotypic MAb were able to enhance the cytolytic activity of a CD8+ T cell clone against ovarian tumour cell lines, demonstrating the presence of functional bi-isotypic MAb (bi-specific MAb). The total amount of immunoglobulin produced after 38 days was 375 mg. After purification by protein A affinity chromatography, 79 mg of bi-isotypic MAb (IgG1/IgG2a) was obtained. This amount of bi-isotypic MAb was similar to the amount obtained from ascitic fluid produced by 200 mice or 38 l of tissue culture flask supernatant.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / immunology
  • Antibody Specificity
  • Cell Line
  • Cytotoxicity, Immunologic
  • Humans
  • Hybridomas
  • Immunoglobulin G / classification
  • Immunoglobulin Isotypes / analysis
  • Mice
  • Mice, Inbred BALB C
  • T-Lymphocytes, Cytotoxic / immunology
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin G
  • Immunoglobulin Isotypes